Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Reaction Pattern of Bacillus cereus D-11 Chitosanase on Chitooligosaccharide Alcohols

  • Gao, Xing-Ai (Glucosamine Saccharide Materials-National Research Laboratory, Division of Applied Bioscience and Biotechnology, Institute of Agricultural Science and Technology, Chonnam National University) ;
  • Jung, Woo-Jin (Glucosamine Saccharide Materials-National Research Laboratory, Division of Applied Bioscience and Biotechnology, Institute of Agricultural Science and Technology, Chonnam National University) ;
  • Kuk, Ju-Hee (Gwangju Regional Korea Food and Drug Administration) ;
  • Park, Ro-Dong (Glucosamine Saccharide Materials-National Research Laboratory, Division of Applied Bioscience and Biotechnology, Institute of Agricultural Science and Technology, Chonnam National University)
  • Published : 2009.04.30
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Abstract

The purified endochitosanase(Mw 41 kDa) from bacterium Bacillus cereus D-11 hydrolyzed chitooligomers $(GlcN)_{5-7}$ into chitobiose, chitotriose, and chitotetraose as the final products. The minimal size of the oligosaccharides for enzymatic hydrolysis was a pentamer. To further investigate the cleavage pattern of this enzyme, chitooligosaccharide alcohols were prepared as substrates and the end products of hydrolysis were analyzed by TLC and HPLC. The chitosanase split $(GlcN)_4GlcNOH$ into $(GlcN)_3+(GlcN)_1GlcNOH$, and $(GlcN)_5GIcNOH$ into $(GlcN)_4+(GlcN)_1GlcNOH$ and $(GlcN)_3+(GlcN)_2GlcNOH$. The heptamer $(GlcN)_6GlcNOH$ was split into $(GlcN)_5$ [thereafter hydrolyzed again into $(GlcN_3+(GlcN)2]+(GlcN)_1GlcNOH$, $(GlcN)_4+(GlcN)_2GlcNOH$, and $(GlcN)_3+(GlcN)_3GlcNOH$, whereas $(GlcN)_{1-3}GlcNOH$ was not hydrolyzed. The monomers GlcN and GIcNOH were never detected from the enzyme reaction. These results suggest that D-11 chitosanase recognizes three glucosamine residues in the minus position and simultaneously two residues in the plus position from the cleavage point.

Keywords

References

  1. Armand, S., H. Tomita, A. Heyraud, C. Gey, T. Watanabe, and B. Henrissat. 1994. Stereochemical course of the hydrolysis reaction catalyzed by chitinases A1 and D from Bacillus circulans WL-12. FEBS Lett. 343: 177-180 https://doi.org/10.1016/0014-5793(94)80314-5
  2. Fenton, D. M. and D. E. Eveleigh. 1981. Purification and mode of action of a chitosanase from Penicillium islandicum. J. Gen. Microbiol. 126: 151-165 https://doi.org/10.1099/00221287-126-1-151
  3. Fukamizo, T., Y. Honda, S. Goto, I. Boucher, and R. Brzezinski. 1995. Reaction mechanism of chitosanase from Streptomyces sp. N174. Biochem. J. 311: 377-383 https://doi.org/10.1042/bj3110377
  4. Gao, X. A., W. T. Ju, W. J. Jung, and R. D. Park. 2008. Purification and characterization of 41-kDa chitosanase from Bacillus cereus D-11. Carbohyd. Polym. 72: 513-520 https://doi.org/10.1016/j.carbpol.2007.09.025
  5. Izume, M., S. Nagae, H. Kawagishi, M. Mitsutomi, and A. Ohtakara. 1992. Action pattern of Bacillus sp. No. 7-M chitosanase on partially N-acetylated chitosan. Biosci. Biotechol. Biochem. 56: 448-453 https://doi.org/10.1271/bbb.56.448
  6. Jo, Y. Y., K. J. Jo, Y. L. Jin, K. Y. Kim, J. H. Shim, Y. W. Kim, and R. D. Park. 2003. Characterization and kinetics of 45 kDa chitosanase from Bacillus sp. P16. Biosci. Biotechnol. Biochem. 67: 1875-1882 https://doi.org/10.1271/bbb.67.1875
  7. Kazuaki, K. and W. Kazuhiro. 2003. Preparation of high purity and colorless chitooligosaccharides and their alcohols. JP Patent 2003-212889A
  8. Lee, H. W., J. W. Choi, D. P. Han, M. J. Park, N. W. Lee, and D. H. Yi. 1996. Purification and characteristics of chitosanase from Bacillus sp. HW-002. J. Microbiol. Biotechnol. 6: 19-25
  9. Lee, Y. S., J. S. Yoo, S. Y. Chung, Y. C. Lee, Y. S. Cho, and Y. L. Choi. 2006. Cloning, purification, and characterization of chitosanase from Bacillus sp. DAU101. Appl. Microbiol. Biotechnol. 73: 113-121 https://doi.org/10.1007/s00253-006-0444-0
  10. Mitsutomi, M., M. Ueda, M. Arai, A. Ando, and T. Watanabe. 1996. Action patterns of microbial chitinases and chitosanases on partially N-acetylated chitosan. Chitin Enzymol. 2: 273-284
  11. Okajima, S., T. Kinouchi, Y. Mikami, and A. Ando. 1995. Purification and some properties of a chitosanase of Nocardioides sp. J. Gen. Appl. Microbiol. 41: 351-357 https://doi.org/10.2323/jgam.41.351
  12. Park, R. D. and Y. L. Jin. 2000. Characteristics, mode of action, and lassification of microbial chitosanases. J. Chitin Chitosan. 5: 207-216
  13. Shimosaka, M., M. Nogawa, X. Y. Wang, M. Kumehara, and M. Okazaki. 1995. Production of two chitosanases from a chitosan-assimilating bacterium, Acinetobacter sp. strain CHB101. Appl. Environ. Microbiol. 61: 438-444
  14. Tremblay, H., T. Yamaguchi, T. Fukamizo, and R. Brzezinski. 2001. Mechanism of chitosanase-oligosaccharide interaction subsite structure of Streptomyces sp. N174 chitosanase and the role of Asp57 carboxylate. J. Biochem. 130: 679-686 https://doi.org/10.1093/oxfordjournals.jbchem.a003034
  15. Tanaka, T., T. Fukui, H. Atomi, and T. Imanaka. 2003. Characterization of an exo-$\beta$-D-glucosaminidase involved in a novel chitinolytic pathway from the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1. J. Bacteriol. 185: 5175- 5181 https://doi.org/10.1128/JB.185.17.5175-5181.2003
  16. Tanaka, T., S. Fujiwara, S. Nishikori, T. Fukui, M. Takagi, and T. Imanaka. 1999. A unique chitinase with dual active sites and triple substrate binding sites from the hyperthermophilic archaeon Pyrococcus kodakaraensis KOD1. Appl. Environ. Microbiol. 65: 5338-5344
  17. Yoon, H. G., H. Y. Kim, H. K. Kim, K. H. Kim, H. J. Hwang, and H. Y. Cho. 1999. Cloning and expression of thermostable chitosanase gene from Bacillus sp. KFB-C108. J. Microbiol. Biotechnol. 9: 631-636

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